Process for preparing water extract of cinnamon

ABSTRACT

This invention relates to an improved process for preparing water extract of cinnamon in a large scale. The process comprises the steps of: (a) adding an aqueous solvent such as water to at least 5 kg of a cinnamon raw material at a water to material ratio of 1:1 to 100:1, (b) boiling the mixture of (a) for at least 5 minutes, (c) removing the solid debris from the mixture, (d) storing the liquid portion of the mixture at about −5 to 25° C., preferably 0-10° C., until a top layer of oil is formed and partitioned, (e) removing the top layer of oil, and (f) collecting the remaining liquid portion. The present process prepares a cinnamon water extract product with a minimal content of potentially toxic cinnamaldehyde and coumarin, while increasing the contents of the active ingredients of polyphenolic polymers for controlling blood glucose level.

This application is a continuation application of U.S. application Ser.No. 12/952,898, filed Nov. 23, 2010; the content of which isincorporated herein by reference in its entirety.

TECHNICAL FIELD

This invention relates to an improved process for preparing waterextract of cinnamon in a large scale. The improved process includes astep of storing the material at a cold temperature to partition the oillayer from the remaining aqueous solution. The improved process reducesthe potentially toxic cinnamaldehyde contents in the final product,while maintaining or increasing the contents of the active ingredientsof cinnamon for controlling blood glucose levels.

BACKGROUND OF THE INVENTION

Diabetes mellitus is a major public health problem. In the UnitedStates, there are over 10 million patients with diabetes. Diabetes is asyndrome that is caused by a relative or an absolute lack of insulin.Clinically, it is characterized by symptomatic glucose intolerance aswell as alterations in lipid and protein metabolism. The maintenance ofnormal blood sugar levels is achieved by the actions of severalhormones, most notably insulin, but also glucagon, epinephrine,corticosteroids, and growth hormone. Hypoglycemia, or low blood sugar,is characterized by below normal levels of blood glucose. On the otherhand, hyperglycemia is exemplified by higher than normal concentrationsof glucose in the blood. The pancreas produces insulin which is releasedin response to increased blood glucose concentrations. Insulin works tolower the blood sugar level by stimulating the uptake of glucose bycells. Glucose is used in cellular metabolism to produce energy, or isconverted to glycogen for storage in the liver and muscles, or is usedin the production of triglycerides and fats.

Water extracts of cinnamon exhibit an insulin potentiating activity,i.e. they increase apparent insulin activity as measured by increasedglucose uptake by cells (U.S. Pat. No. 6,200,569). Improved insulinactivity leads to decreased circulating insulin, which leads to lowerblood glucose and lower glycosylated hemoglobin levels in patients; italso has an effect on smoothing out fluctuations in glucose levels. Inthe '569 patent, water extracts were prepared by extracting cinnamonplants with water, removing the solid debris, and collecting the liquidextracts. The liquid extracts could be used directly or dried to apowder form.

Mang et al (European J. Clin. Invest. 36:340-344, 2006) report thatdiabetes mellitus type 2 patients treated with 3 g of cinnamon waterextract powder per day for 4 months decreased fasting plasma glucoselevel, in comparison with placebo-treated patients.

Anderson et al (J. Agric. Food Chem. 52:65-70, 2004) report theisolation and characterization of polyphenol type-A polymers fromcinnamon. Two trimers with a molecular mass of 864 and a tetramer with amass of 1152 were isolated. These polymers are water-soluble and arebelieved to be the active ingredients of cinnamon that potentiateinsulin action and control glucose metabolism.

Cinnamon is composed of between 1-8% of essential oils, in which 65-90%is cinnamaldehyde. (BfR (Bundesinstitut für Risikobewertung) HealthAssessment No. 044/2006, Aug. 18, 2006). He et al (J. Agric Food Chem.53:2424-2428, 2005) found that in 1 g of cassia cinnamon (raw material),cinnamaldehyde level was between 13.1 and 56.9 mg (mean 28.9 mg).Cinnamaldehyde is a yellowish oily liquid and solidifies at −7.5° C.(Merck Index).

Lewis et al. (Environ Health Perspect 104:1011-1016, 1996)) report thatcinnamaldehyde was tested positive for potential carcinogenicity in theCOMPACT evaluation.

The chemical name of coumarin is 1-benzopyran-2-one (CAS Number:91-64-5). Coumarin is poorly soluble in water but readily soluble inalcohol. Coumarin is a naturally occurring flavoring substance which iscontained in many plants such as woodruff, sweet clover, Tonka beans.Higher concentrations can be found particularly in some types ofcinnamon. Large amounts of coumarin can cause hepatic damage—coumarinmay only be used as an ingredient of flavorings and other food additiveswith flavoring properties in the food sector. In animal experiments,coumarin can trigger the formation of tumors. (see BfR Health AssessmentNo. 043/2006, 16 Jun. 2006) Coumarin is not as potentially toxic ascinnamaldehyde. Still, it is undesirable to have large amounts ofcoumarin for human consumption.

There exists a need for an improved process to prepare water extract ofcinnamon. The improved process reduces the contents of potentially toxiccinnamaldehyde and coumarin in the final product, while maintaining orincreasing the active ingredients of cinnamon.

SUMMARY OF INVENTION

This invention is directed to an improved process for preparing waterextract of cinnamon in a large scale. The process comprises the stepsof: (a) adding an aqueous solvent such as water to a cinnamon rawmaterial at a solvent to material ratio of 1:1 to 100:1 (w/w), (b)boiling the mixture for at least 5 minutes, (c) removing the soliddebris from the mixture to obtain a liquid portion, (d) storing theliquid portion of the mixture at −5 to 25° C. until a top oil layer isformed and partitioned, (e) removing the top oil layer, and (f)collecting the remaining liquid portion. After the step (f), the processoptionally comprises a step (g) of centrifuging the remaining liquidportion with a tubular centrifuge to remove solids of less than 1 micronand collect the supernatant.

The present process is suitable for processing 5-5,000 Kg of a cinnamonraw material. The present process prepares a cinnamon water extractproduct with a reduced amount of potentially toxic cinnamaldehyde, whileincreasing the contents of the active ingredients of polyphenolicpolymers for controlling blood glucose level.

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to an improved process for preparingcinnamon water extract. The inventors have discovered that by storingthe liquid water extract of cinnamon at a cold temperature (−5 to 25°C., preferably 0-10° C.), a layer of oil is formed and separated fromthe remaining aqueous portion. By removing the oil layer, thecinnamaldehyde content in the final product is reduced, whereas thecontents of the active ingredients that are effective to potentialinsulin activity and to lower blood glucose level are increased.

The process of the present invention comprises the steps of: (a) addingan aqueous solvent such as water to a cinnamon raw material at a waterto material ratio of 1:1 to 100:1 (w/w), (b) boiling the mixture for atleast 5 minutes, (c) removing the solid debris from the mixture toobtain a liquid portion, (d) storing the liquid portion of the mixtureat −5 to 25° C. until a top oil layer is formed and partitioned, (e)removing the top oil layer, and (f) collecting the remaining liquidportion.

After the step (f), the process optionally comprises a step (g) ofcentrifuging the remaining liquid portion with a tubular centrifuge toremove small solids and collect the supernatant. The supernatant isoptionally dried to obtain the cinnamon water extract in a dry powderform.

Alternatively, after step (f), the process optionally comprises a stepof drying the liquid portion of (f) to obtain the cinnamon water extractin a dry powder form.

The process of the present invention is particularly suitable for alarge scale production. Large scale, as used herein, refers to thestarting material of about ≧5 Kg or ≧10 Kg, preferably ≧50 Kg,preferably ≧100 Kg, more preferably ≧500 Kg, and most preferably ≧1000Kg. The present invention is suitable for processing a startingmaterial, for example, in the range of about 5-2,000 Kg, 10-2,000 Kg,10-5,000 Kg, 50-2,000 Kg, 50-5,000 Kg, 100-2,000 Kg, 100-5,000 Kg,400-2,000 Kg, 400-5,000 Kg, 1,000-5,000 Kg, or 1,000-10,000 Kg.

“About” as used in this application, refers to ±15% of the recitedvalue.

Preferred source of a raw material of cinnamon is bark from a cinnamontree, in the family of Cinnamomum. Preferred species are Cinnamomummairei, Cinnamomum zeylanicum, Cinnamomum burmannii, and Cinnamomumcassia. Cinnamomum mairei is a tree with highly aromatic bark, whichbark can be used for preparing extracts. Commercial cinnamon bark, whichis the dried inner bark of the shoots, and ground cinnamon obtained froma grocery store can also be used for preparing extracts.

In the process step (a), the cinnamon raw material is obtained either asa ground powder or is prepared by cutting the plant into small pieces,pulverizing it, grinding it or otherwise increasing the surface area ofthe pieces of tissue to facilitate extraction.

In the process step (a), aqueous hydrophilic solvents are used forextraction of active ingredients that have insulin potential activity.Because it is safe, easy to use, and economic, water (de-ionized water,distill water, purified water, or tap water) is a preferred solvent forextraction. In addition, water does not extract those impurities solubleonly in acid or base. A small amount of buffer can be added to the waterto maintain the pH. A small amount of acid, base, ethanol, or methanolalso can be added to the distilled water as a solvent for extraction.However, high concentrations, such as 50%, of ethanol can extractundesired organic impurities such as cinnamaldehyde, which is toxic whenpresent in large amounts for human consumption. In the process step (a),the aqueous solvent to the raw material ratio is about 1:1, or 2:1, or5:1, or 10:1, or 20:1, or 30:1, or 100:1 (w/w); or any range in betweenthe above recited ratios.

In the process step (b), the water/raw material mixture is boiled for atleast 5 minutes, or 10 minutes, or 30 minutes, or an hour, or 2 hours,or 4 hours, or 6 hours, or 8 hours, or 10 hours, or any range in betweenthe above recited time. The desired boiling time depends on the amountof the raw material and water. The desired boiling time for a largeamount of raw materials is longer than that for a small amount of rawmaterials. The process steps (a)-(c) can optionally be repeated severaltimes, e.g., 2-8 times, or 2-4 times. After each boiling step, thesupernatant is removed and collected, and new water is added to thesolid material, and the mixture is boiled. After the boiling steps arecompleted, the supernatants are combined.

In the process step of (c), the liquid portion is separated from thesolid debris by centrifugation, filtration, or decanting. The soliddebris is discarded, and the liquid portion is collected.

The liquid portion is optionally concentrated to a smaller volume forconvenient handling. Typically, the liquid portion is concentrated by5-50 folds, and the volume is typically reduced to less than 1000 L. Theconcentration can be carried out by any means, for example, evaporation,vacuum evaporation, or other means known to a skilled person forconcentration of a liquid material.

Then the liquid portion is stored at a cold temperature for a sufficienttime until a top layer of oil is formed and partitioned from theremaining aqueous liquid portion. The temperature and time of coldstorage depends on the volume of the liquid and the size of thecontainer. When the liquid portion is cold, the oil layer forms; the oillayer is immicible with the aqueous liquid and floats to the top. Ingeneral, the temperature is from about −5 to 25° C., preferably about0-25° C., and more preferably about 0-20° C., 0-15° C., 0-10° C., or0-5° C. A small fluctuation of temperature, i.e. ±2° C., during the coldstorage, is included in this invention. The cooler the temperature, thesmaller amount of the liquid, the less time it takes to have the oillayer formed. In general, the liquid portion is stored at a coldtemperate for at least one hour, or two hours, or four hours, or 8hours, or overnight, or 24 hours, or 48 hours, or any range in betweenthe above-recited time. After the cold storage, the oil layer partitionsout from the aqueous liquid and floats to the top.

The undesired potentially toxic material, cinnamaldehyde, which islipid-soluble, is portioned to the top oil layer. The top oil layer isremoved carefully by withdrawing or decanting. The toping layer is thendiscarded. By removing the oil layer, the cinnamaldehyde content in theremaining aqueous liquid is reduced 2-6 fold.

The oil-removal step does not reduce the contents of active ingredients,i.e., polyphenol type-A polymers, because polyphenolic polymers arewater-soluble and not lipid-soluble (Anderson et al. J. Agric. FoodChem. 52:65-70, 2004). The inventors have discovered that theoil-removal step increases the percent content of polyphenolic polymersin the final product.

After the oil-layer is removed, the remaining aqueous liquid isoptionally centrifuged in a tubular centrifuge to remove suspendedsolids which are between 0.001-1 microns in size. Tubular centrifugesoperate at high speed, generating up to 20,000×g, causing liquid todischarge through the outlet and solids to accumulate inside thecentrifuge. This tubular centrifugation step is typically carried out atabout 14,000-20,000 RPM (about 15,000-17,000×g) at a flow rate of about20-1200 L/hour, depending on the viscosity of the liquid. This tubularcentrifugation step significantly reduces the content of coumarin, whichis an undesirable material, in the final product. This tubularcentrifugation step also further reduces the content of cinnamaldehyde.

The remaining aqueous liquid can be used directly as an insulinpotentiating agent, or the aqueous liquid can be dried to form a powder.The drying can be carried out by any means that can practically dry alarge quantity of liquid (over 1 kg, 10 kg, or 100 kg), e.g.,lyophilization, spray-drying, or centrifuging and spray-drying. When theoptional tubular centrifugation step is omitted in the process, theyield of the dry powder is about 5-20% and often about 8-12% (e.g. about10%) of the weight of the starting raw material. When the optionaltubular centrifugation step is included in the process, the yield of thedry powder is about 3-18% and often about 5-11% of the weight of thestarting raw material. The tubular centrifugation step improves thequality of the product, while it may decrease the yield. Depending onthe intended use of the final product, the tubular centrifugation stepis either included or excluded from the process of this invention.

The process of the present invention improves the water solubility ofthe product from about 92% to 99%, which is particularly important whenthe product is used as a beverage. For a beverage, it is desirable tohave a minimal amount (≦1%) of insoluble materials. Otherwise, thebeverage may not be commercially viable because consumers do not like totaste solids when drinking a beverage.

The liquid or the powder can be incorporated into a variety of basicmaterials in the form of a liquid, powder, tablets, or capsules to givean insulin potentiating activity effective to decrease blood glucoselevels or glycosylated hemoglobin levels. Such a product is effective inlowering blood glucose and glycosylated hemoglobin levels, and iseffective in the treatment of hyperglycemia.

EXAMPLES Example 1 Preparation of Water Extract of Cinnamon

-   1. 1,200 kg of raw materials of cinnamon bark (Cinnamomum Cassia)    was divided among 4 tanks, each holding 300 kg of the raw material.-   2. Purified water was added into each tank at water to material    ratio of 8:1, and the mixture was heated to boil at 100° C. for one    hour. The first water extract was removed, and more water was added    to the cinnamon solid material at water to material ratio of 6:1;    the mixture was boiled at 100° C. for another hour. The second water    extract was removed, and more water was added to the cinnamon solid    material at water to material ratio of 4:1; the mixture was boiled    at 100° C. for another hour. The third water extract was removed.

The water extracts were combined and filtered through a 100-mesh screento remove solid debris and the filtrates were collected.

-   3. The filtrates were put into intermediate containers and held    static for 4 hours. After static holding, the supernatants were    carefully removed and put into a concentration tank for vacuum    evaporation to concentrate the filtrates.-   4. The vacuum evaporation was performed at 60° C. to 80° C., with    vacuum pressure from −0.06 MPa to −0.08 MPa. After 24 hours, a total    of 600 L concentrate was obtained.-   5. The concentrate was stored in a container at 0-5° C. for 16-24    hours and a top oil layer was formed and separated out from the    remaining concentrate. The top oil layer (20-60 L) was removed and    discarded.-   6. After the top oil layer was removed, the concentrate was    optionally centrifuged in a tubular centrifuge (Model GQ 105,    Shanghai Centrifuge Institute Co., Ltd.) at 16,000 RPM at about 2    L/minute flow rate. The precipitate was removed and about 500 L of    liquid collected. This tubular centrifugation step was optional.-   7. The liquid from step 5 or step 6 was then sprayed dry in a    high-speed centrifuging and spray-drier (Model: LHS25, Menghe Drying    Equipment Works, Jiangsu, China) to form dry powder. The dry powder    was mixed and the yield of cinnamon water extract in the dry powder    form was about 100-120 kg without step 6, and about 50-110 Kg with    step 6, depending on the batch.

Example 2 Comparison of the Contents of Cinnamaldehyde, Coumarin, andActive Ingredients in Products by Three Different Processes

Objectives

The contents of cinnamaldehyde, coumarin, and active ingredients(polyphenolic polymers, see Anderson et al, J. Agric. Food Chem.52:65-70, 2004) in the dry powder of water extracts of cinnamon preparedby three different processes were compared.

The water solubility of the dry powder of water extracts of cinnamonprepared by three different processes was also compared.

Process 1 (prior method): dry powder of water extract of cinnamon wasprepared by the similar protocols of Example 1, except Step 5 (coldstorage and oil removal) and step 6 (centrifugation) were not performed.

Process 2 (invention): dry powder of water extract of cinnamon wasprepared by the protocols of Example 1, without the optional step 6(centrifugation).

Process 3 (invention): dry powder of water extract of cinnamon wasprepared by the protocols of Example 1, including the optional step 6(centrifugation).

Methods

Polyphenolic Polymers and Coumarin

Material Preparation for Testing: 10 mg of dry powder was added to 1 mLof 0.1 N acetic acid. The sample was then centrifuged to removeinsoluble and the dry weight of the solution was determined (mg/ml). 0.6mL of the solution was added to Prep C18 column.

Standards: Coumarin standards were purchased from Sigma Aldrich. Thestandards of polyphenolic polymers were determined by mass spectroscopy(Anderson et al, 2004).

HPLC Column: 7.8×300 mm Symmetry Prep C18 column, (Waters WAT 066235);0.1N Acetic acid 92%, 8% acetonitrile for 90 min; gradient to 15%acetonitrile at 130 min; constant to 160 min; gradient to 20%acetonitrile at 180 min; at 200 min, run 8 min gradient to 100%acetonitrile. The absorbance area at 278 nm of each peak was determinedand compared with that of a standard. The amount and the identity of thepeaks were determined based on the comparison with the standards.

Content Calculation: The amounts of coumarin, and polyphenolic polymerswere determined based on the comparison with the standards. Eachdetermined amount was divided by the amount of material added to thecolumn and then converted to percent.

Cinnamaldehyde

Cinnamaldehyde was determined according the method described in ChinesePharmacopoeia 2005, page 91.

Material Preparation for Testing: 0.5 g of the dry power was weighed.The dry powder was put in a conical flask and 25 ml of methanol wasadded. The mixture was mixed with an ultrasonic device for 10 minutes(power 350 W, frequency 35 kHz), then stored overnight. The mixture wasmixed again with the ultrasonic device for 10 minutes and the weight wasmeasured. Methanol was added to supply the lost weight. The solution wasthoroughly mixed and then filtered. Exactly 1 ml of the filtrate wasremoved, added with 24 mL of methanol, and mixed.

Standard Preparation: Cinnamaldehyde (Sigma Aldrich) was weighed andadded with a proper amount of methanol, to prepare the concentration of10 μg/mL.

HPLC Column: 10 μL of test sample or standard solution was injected intoPhenomenex Luna C18 column. The mobile phase was acetonitrile-water(35:75 v/v) and the test wavelength was 290 nm.

Content Calculation: The amount of cinnamaldehyde in the test sample wasdetermined based on the comparison with the standard. The determinedamount was divided by the amount of material added to the column andthen converted to percent.

Water Solubility

0.1 g of dry powder was added with 100 mL of warm water (about 60° C.),and mixed by an untrasonic device for 30 minutes. The mixture was keptat about 60° C. and filtered through a G3 filter funnel. The filtratewas dried and weighed.Water-solubility (%)=(1−dry filtrate weight/sample weight)×100%Results

The results are summarized in Table 1.

TABLE 1 The percent contents of cinnamaldehyde, coumarin, andpolyphenolic polymers in the dry powder and the water solubility of thedry powder prepared by three different processes. Coumarin PolyphenolSolubility Cinnamaldehyde Content Polymers in Water Content (%) (%)Content (%) (%) Process 1 0.006 0.66 3.52 91.84 (Prior Method) Process 20.001 0.60 5.20 99.19 (Invention) Process 3 0.0005 0.29 4.20 99.53(Invention)

The potentially toxic cinnamaldehyde content in the process 1 product(prior art) was 0.006%, and the cinnamaldehyde contents in the process 2and process 3 products were 0.001 and 0.0005%, respectively. The presentprocesses reduced the cinnamaldehyde content by at least 6-fold.

The potentially toxic coumarin content in the process 1 product (priorart) was 0.66%, and the coumarin contents in the process 2 and process 3products were 0.60 and 0.29%, respectively. The present process 3 withthe centrifugation step reduced the coumarin contents by more than 2fold; this could be due to the low solubility of coumarin in water.

The polyphenol polymer content in the process 1 product (prior art) was3.52%, and the polyphenol polymer contents in the process 2 and process3 products were 5.20 and 4.20%, respectively. The present processesincreased the percent content of the active ingredients in the finalproducts.

The present processes 2 and 3 improve the water solubility of theproduct from about 92% to over 99%, which is particularly important whenthe product is used as a beverage. For a beverage, it is desirable tohave a minimal amount (≦1%) of insoluble materials.

The invention, and the manner and process of making and using it, arenow described in such full, clear, concise and exact terms as to enableany person skilled in the art to which it pertains, to make and use thesame. It is to be understood that the foregoing describes preferredembodiments of the present invention and that modifications may be madetherein without departing from the scope of the present invention as setforth in the claims. To particularly point out and distinctly claim thesubject matter regarded as invention, the following claims conclude thisspecification.

1. A process for preparing cinnamon water extract, comprising the stepsof: (a) adding an aqueous solvent to at least 5 Kg of a cinnamon rawmaterial, wherein the ratio of the solvent to the material is 1:1 to100:1 (w/w), (b) boiling the mixture of (a) for at least 5 minutes, (c)removing the solid debris from the mixture to obtain a liquid portion,(d) concentrating the liquid portion by 5-50 fold, (e) storing theconcentrated liquid portion at about −5° C. to 25° C. until a top oillayer is formed and partitioned, (f) removing the top oil layer, and (g)collecting the remaining liquid portion.
 2. The process according toclaim 1, wherein the aqueous solvent is water.
 3. The process accordingto claim 2, after the step (g), further comprising a step (h) ofcentrifuging the remaining liquid portion by a tubular centrifuge toremove suspending solids and collecting the supernatant.
 4. The processaccording to claim 3, further comprising a step of drying thesupernatant of (h) to obtain the cinnamon water extract in a dry powderform.
 5. The process according to claim 2, further comprising a step ofdrying the liquid portion of (g) to obtain the cinnamon water extract ina dry powder form.
 6. The process according to claim 1, wherein theconcentrating is performed by vacuum evaporation.
 7. The processaccording to claim 1, further comprising repeating steps (a)-(c) 2-4times with the same cinnamon raw material, wherein after each boilingstep, the liquid portion is removed and collected, and new water isadded to the solid material for boiling.
 8. The process according toclaim 1, wherein the ratio of the solvent to the cinnamon raw materialis 2:1 to 50:1 (w/w).
 9. The process according to claim 8, wherein theratio is 10:1 to 30:1 (w/w).
 10. The process according to claim 1,wherein the mixture of (a) is boiled for 5 minutes to 10 hours.
 11. Theprocess according to claim 10, wherein the mixture of (a) is boiled for1-4 hours.
 12. The process according to claim 1, wherein theconcentrated liquid portion of (e) is stored at 0-10° C. for at least 1hour.
 13. The process according to claim 12, wherein the concentratedliquid portion of (e) is stored at 0-10° C. for 4-48 hours.
 14. Theprocess according to claim 1, wherein the cinnamon raw material is atleast 100 Kg.
 15. The process according to claim 14, wherein thecinnamon raw material is at least 1000 Kg.
 16. The process according toclaim 15, wherein the cinnamon raw material is 100-2000 Kg.
 17. Theprocess according to claim 4, wherein the cinnamon raw material is atleast 100 Kg, the mixture of (a) is boiled for 1-4 hours, and theconcentrated liquid portion of (e) is stored at 0-10° C. for 4-48 hours.18. The process according to claim 5, wherein the cinnamon raw materialis at least 100 Kg, the mixture of (a) is boiled for 1-4 hours, and theconcentrated liquid portion of (e) is stored at 0-10° C. for 4-48 hours.19. The process according to claim 1, wherein the cinnamon raw materialis bark from Cinnamomum mairei, Cinnamomum zeylanicum, Cinnamomumburmannii, and Cinnamomum cassia.